HEATING BLOCK

MICRO STRIP READER

MECHANICAL WASHER

HEATING BLOCK

MICROSTRIP READER

MECHANICAL WASHER

PRINCIPLE OF ELISA ELISA (ENZYME-LINKED IMMUNOSORBENT ASSAY) is a widely used biochemical technique for the detection of an antigen in a sample. The sandwich ELISA utilizes two antigen specific antibodies, a capture antibody bound to a solid phase and an enzyme linked detection antibody. Direct enzyme conjugation of the detection antibody ensures an easy-to-use and sensitive assay with minimal background signal. Sandwich ELISA sequence assay - antigen and detection antibody is added stepwise

In a sequence assay, sample is applied to an antibody-coated microtiter plate well. The capture antibody binds to the antigen in the sample. Unbound compounds are removed by a washing procedure before the enzyme-linked detection antibody is added to the well. The detection antibody binds to a second epitope (immunogenic part) on the antigen. Unbound antibodies are removed by washing before the addition of a substrate, which is converted by the enzyme to a chromogenic signal. The enzyme reaction is stopped and the result is monitored spectrophotometrically. The more antigen in the sample, the stronger the signal. The sandwich ELISA may also be constructed as a simultaneous assay where the sample and detection antibody is added in the same step.

Sandwich ELISA simultaneous assay - antigen and detection antibody is added simultaneously

Competitive ELISA assay

A second type of ELISA is the competitive assay in which there is a competitive binding of an antigen-specific, biotin-linked antibody to sample antigen or to antigen bound to the microtiter well. Bound antibody is detected with enzyme-linked streptavidin. Because the concentration of antigenspecific antibody is important and must be limited in this assay, a biotin/streptavidin step is used to enhance the signal. In this assay, the more antigen in the sample, the weaker the signal. Several alternative variants of the competitive ELISA format are found. Reference: http://www.mercodia.se/learning-center/mercodia-elisa-technology/principle-of-technology.html

BASIC PRINCIPLE OF ELISA Use an enzyme to detect the binding of antigen (Ag) antibody (Ab). The enzyme converts a colorless substrate (chromogen) to a colored product, indicating the presence of Ag : Ab binding. An ELISA can be used to detect either the presence of Antigens or antibodies in a sample depending how the test is designed.

IMMUNOASSAY FORMATS